Interaction of the fluorescent probe N-(lissamine Rhodamine B sulfonyl)dipalmitoylphosphatidylethanolamine with phosphatidylcholine bilayers.

The surface density of the fluorescent probe N-(lissamine Rhodamine B sulfonyl)dipalmitoylphosphatidylcholine is the same in the two lipid leaflets of phosphatidylcholine bilayers containing the probe. In the liquid-crystalline state, the probe molecules aggregate above a threshold amount, approximately 0.2 mol/mol phospholipids. Above this threshold value, the surface density of the free probe molecules is constant, and all probe molecules added are incorporated in the aggregated form. The aggregation of the probe increases by approximately 20% when the medium pH is lowered to 4. In the gel state, the probe aggregation is higher than that in the liquid-crystalline state, and the free probe molecules distribute unevenly in the bilayer surface. Even though the results obtained in our model system cannot be directly extrapolated to all model systems, we point out that care is to be taken in the use of the probe. In fact, only in membranes in the liquid-crystalline state in which the amount of probe molecules to phospholipid molecules is lower than 1:7 the fluorescence response of the probe is independent of the pH changes and of the molecular aggregation.